Journal: Neurobiology of disease

Article Title: Inhibition of CXCR2 enhances CNS remyelination via modulating PDE10A/cAMP signaling pathway.

doi: 10.1016/j.nbd.2023.105988

Figure Lengend Snippet: Fig. 3. PDE10A regulated differentiation and maturation in primary cultured OPCs. Primary OPCs were maintained for 5 days in differentiation medium containing 0.1 μM and 1 μM TAK-063 or DMSO as control. (A) Confocal results of A2B5 expression in primary cultured OPCs. (B) Confocal results of MBP expression in primary cultured OPCs. Cells were stained with A2B5 (green), MBP (red) and DAPI (blue). (Scale bar = 50 μm). (C, D) Western blot bands and statistical analysis of PDGFRα, MBP, PLP1, MOG and MOBP expression. Primary cultured OPCs were transfected with PDE10A or control cDNA for 6 h, and then incubated with SB225002 at 1 μM for another 24 h. (E) Western blot bands of PDGFRα and MBP in PDE10A overexpressed cells. (F, G) Relative value of PDGFRα and MBP intensity normalized by β-actin. Results are shown as the mean ± SD form at least five experiments, and the confocal assay was performed three independent times with duplication. *P < 0.05, **P < 0.01 and ***P < 0.001 vs. Control cells, &&&P < 0.01 vs. OPC overexpressed PDE10A. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The blots were incubated with primary antibodies overnight at 4 ◦C, including CXCR2 (A3301), PLP1 (A14251), MAG (A16914), MOG (A5353), Caspr (A3721), SOX10 (A8655) (1:1000), β-actin (AC026, 1:100000) (Abclonal, China), MOBP (12472–1-AP) (1:500, Proteintech, China), PDE10A (sc-515,023), PDGFRα (sc-398,206) (1:500, Santa Cruze, USA), MBP (Ab40390), OLIG2 (Ab109186) (1:1000, Abcam, USA), pERK1/2 (4695 s), ERK1/2 (4370 s), Lamin B1 (13435 s) (1:1000, Cell signaling technology, USA).

Techniques: Cell Culture, Control, Expressing, Staining, Western Blot, Transfection, Incubation, Confocal Assay

Journal: Neurobiology of disease

Article Title: Inhibition of CXCR2 enhances CNS remyelination via modulating PDE10A/cAMP signaling pathway.

doi: 10.1016/j.nbd.2023.105988

Figure Lengend Snippet: Fig. 4. Inhibition of PDE10A promoted remyelination in EB intoxicated rats. MS model rats were induced with EB injection into right lateral ventricle and then treated with TAK-063 (2 mg/kg) for 14 days. (A) Rotarod test of rats treated with TAK-063 (n = 10). (B, D) LFB staining and statistical analysis of demyelinated area in the rat corpus callosum. (C, E) Expression and calculation of the mean IOD of MBP in the rat corpus callosum. (F) Western blot assay of PDGFRα, MBP, MOG, MOBP and Caspr expression in the rat corpus callosum. (G) Relative value of PDGFRα, MBP, MOG, MOBP and Caspr intensity normalized by β-actin. Results are presented as the mean ± SD from at least four rats. **P < 0.01 and ***P < 0.001 vs. Control rats, ##P < 0.01 and ###P < 0.001 vs. EB-injected rats.

Article Snippet: The blots were incubated with primary antibodies overnight at 4 ◦C, including CXCR2 (A3301), PLP1 (A14251), MAG (A16914), MOG (A5353), Caspr (A3721), SOX10 (A8655) (1:1000), β-actin (AC026, 1:100000) (Abclonal, China), MOBP (12472–1-AP) (1:500, Proteintech, China), PDE10A (sc-515,023), PDGFRα (sc-398,206) (1:500, Santa Cruze, USA), MBP (Ab40390), OLIG2 (Ab109186) (1:1000, Abcam, USA), pERK1/2 (4695 s), ERK1/2 (4370 s), Lamin B1 (13435 s) (1:1000, Cell signaling technology, USA).

Techniques: Inhibition, Injection, Staining, Expressing, Western Blot, Control

Journal: Autophagy

Article Title: Microglial autophagy defect causes parkinson disease-like symptoms by accelerating inflammasome activation in mice.

doi: 10.1080/15548627.2020.1719723

Figure Lengend Snippet: Figure 3. PDE10A is involved in the activation of NLRP3 inflammasome. (A) The model of LC-MS/MS analysis. (B) The data shows the numbers of upregulated and downregulated genes in 3-month and 8-month WT and Atg5 cKO mice. (C) Immunoblotting analysis of PDE10A and GAPDH in the striatum from 3-month and 8-month-old Atg5 WT mice and Atg5 cKO mice as indicated. (D) Quantitative data of PDE10A levels in the striatum in 3-month and 8-month-old Atg5 WT and Atg5 cKO mice. (E) Immunoblotting analysis of PDE10A, LC3 and GAPDH in the primary microglial cells treated with Baf A1 (50 nM, 6 h), MG132 (10 µM, 6 h) and Rapamycin (20 nM, 6 h). Experiments were carried out for 3 independent times. (F) Primary microglial cells were treated with Baf A1 (50 nM, 6 h). Cell lysates were immunoprecipitated with anti-PDE10A antibody coated gel and then blotted for Ubiquitin, PDE10A, SQSTM1, LC3 and GAPDH. (G) Immunofluorescent staining of cAMP in the primary microglial cells treated with or without Baf A1 (50 nM, 6 h). Scale bar represents 10 µM. N = 50. (H) Quantitative data of cAMP levels as indicated. (I) Immunoblotting analysis of cleaved CASP1 and IL1B levels in the supernatant, Pro-IL1B, total CASP1 and ACTB levels in the pellets from the primary microglial cells treated with Baf A1 (50 nM, 6 h) and MP-10 (5 µM, pre-treatment for 1 h) as indicated. Experiments were carried out 3 independent times. (J) Quantitative data of IL1B levels as indicated. (K) Immunoblotting analysis of cleaved CASP1, total CASP1 and GAPDH in the striatum from 3-month-old Atg5 WT and Atg5 cKO mice as shown. (L) Quantitative data of cleaved CASP1 levels in the striatum in 3-month-old Atg5 WT and Atg5 cKO mice. *means p < 0.05. (M) Immunoblotting analysis of PDE10A, NLRP3, cleaved CASP1, total CASP1 and GAPDH in the SN from AAV-shvector injection group mice and AAV-shAtg5 injection group mice as indicated. (N-P) Quantitative data of PDE10A, NLRP3 and cleaved CASP1 levels. *means p < 0.05. ***means p < 0.001.

Article Snippet: Immunoblots were probed with the following primary antibodies and visualized by ECL (Thermo Fisher, 32106): NLRP3 (Adipogen, AG-20B-0014; 1:1000), CASP1/caspase-1 (Adipogen, Ag-20B-0042; 1:1000), IL1B/IL1β (R&D, AF401; 1:1000), SNCA/ α-synuclein (Abcam, ab1903; 1:1000), PDE10A (Santa Cruz Biotechnology, sc-515023; 1:1000), MIF (Abclonal, A1391; 1:1000), APOE (Abclonal, A10963; 1:1000), LC3 (Cell Signaling Technology, 2775; 1:1000), SQSTM1 (MBL, PM045; 1:1000), ATG5 (MBL, M153-3; 1:1000), Ubiquitin (Cell Signaling Technology, 3933; 1:1000), GAPDH (Cwbiotech, CW0100A; 1:1000).

Techniques: Activation Assay, Liquid Chromatography with Mass Spectroscopy, Western Blot, Immunoprecipitation, Ubiquitin Proteomics, Staining, Injection

Journal: Cancer Medicine

Article Title: Genomic characterization of individuals presenting extreme phenotypes of high and low risk to develop tobacco‐induced lung cancer

doi: 10.1002/cam4.1500

Figure Lengend Snippet: Univariate and multivariate Cox regression analyses of PDE10A protein expression for recurrence‐free survival (RFS) and overall survival (OS) in patients with stage I–II NSCLC

Article Snippet: We used an anti‐human PDE10A antibody (Genetex) diluted 1:500.

Techniques: Expressing